p-Smad3 differentially regulates the cytological behavior of osteoclasts before and after osteoblasts maturation.

BACKGROUND: A series of previous investigations have revealed that p-Smad3 plays a facilitative role in the differentiation and maturation of osteoblasts, while also regulating the expression of certain intercellular communication factors. However, the effects of p-Smad3 in osteoblasts before and after maturation on the proliferation, migration, differentiation, apoptosis and other cellular behaviors of osteoclasts have not been reported. METHODS: MC3T3-E1 cells were cultured in osteogenic induction medium for varying durations, After that, the corresponding conditioned medium was collected and the osteoclast lineage cells were treated. To elucidate the regulatory role of p-Smad3 within osteoblasts, we applied the activator TGF-ß1 and inhibitor SIS3 to immature and mature osteoblasts and collected corresponding conditioned media for osteoclast intervention. RESULTS: We observed an elevation of p-Smad3 and Smad3 during the early stage of osteoblast differentiation, followed by a decline in the later stage. we discovered that as osteoblasts mature, their conditioned media inhibit osteoclasts differentiation and the osteoclast-coupled osteogenic effect. However, it promotes apoptosis in osteoclasts and the angiogenesis coupled with osteoclasts. p-Smad3 in immature osteoblasts, through paracrine effects, promotes the migration, differentiation, and osteoclast-coupled osteogenic effects of osteoclast lineage cells. For mature osteoblasts, p-Smad3 facilitates osteoclast apoptosis and the angiogenesis coupled with osteoclasts. CONCLUSIONS: As pre-osteoblasts undergo maturation, p-Smad3 mediated a paracrine effect that transitions osteoclast cellular behaviors from inducing differentiation and stimulating bone formation to promoting apoptosis and coupling angiogenesis.

Synthesis of a Quaternary Ammonium-Halamine and Preparation on the Modified Nanofibrous Filter with Superior Sterilization, Air Filtration, and Biodegradability.

The outbreak of the 2019 coronavirus pandemic has raised worldwide attention about self-protection from airborne diseases. Air filtration and wearing mask have been proven to be effective measures in reducing the pathogenic aerosol's transmission. Those lead to an increasing demand of high-efficient filters. However, the nonbiodegradable polymeric materials used in filters can accumulate in landfills or ecosystems, potentially causing pollution after improper disposals. Sustainable and biodegradable alternatives to current filter materials are urgently needed. Yet, very few commercial filters meet these needs. In this paper, a novel quaternary ammonium-halamine compound containing Schiff base and a sandwich-structured preparation strategy were developed. The obtained multifunctional filter consists of a PLA fleece as a support layer, an antimicrobial coating for bactericidal function, and a nanofibrous membrane for the particle removal. The filter demonstrates strong bactericidal properties, killing 97% of Escherichia coli and Staphylococcus aureus at a biocide concentration of only 1 mg/mL. It can rapidly kill bacteria within 5 min contact without leaching antimicrobial substances. Furthermore, it boasts a filtration performance with a success rate over 99.99% and a pressure drop of 45 Pa, which surpasses that of commercial N95 filters for PM0.3. Even under humid conditions, it maintains excellent filtration performance. Our reusability testing result of the developed filters shows that a simple halogenation treatment can renew the halamines and restore the filter's antimicrobial activity. The filters can degrade in natural soil. The successful development of this sustainable and biodegradable filter material offers a new alternative for high-performance air quality control that protect public health.

The Mechanism of Action and Experimental Verification of Narenmandula in the Treatment of Postmenopausal Osteoporosis.

BACKGROUND: Narenmandula is a classic ancient remedy in Inner Mongolia, historically used for gastrointestinal diseases. In recent decades, Inner Mongolia Medical University found that it has a significant effect in promoting fracture healing and increasing bone density, and has been used to treat postmenopausal osteoporosis (PMOP), but its mechanism is unclear. OBJECTIVE: Identify the mechanism of action of Narenmandula for PMOP treatment. METHODS: Network pharmacology, molecular docking and ovarian departing rat models were used to verify the relevant mechanism of Narenmandula in the treatment of PMOP. RESULTS: We confirmed that NRMDL prescription can improve OVX-induced bone loss, improve trabecular density, and relieve osteoporosis. Upon screening of network pharmacology, we obtained 238 overlapping genes of Narenmandula and PMOP, and analyzed AKT, IL1B, and IL6 as key genes by network topology. Among the 1143 target genes that interact with PMOP, 107 NRMDL active compounds correspond to 345 target genes and 238 overlapping genes. Network topology analysis showed the top 8 active ingredients, such as quercetin and kaempferol, and the top 20 key genes, such as AKT, IL1B, IL6, INS, JUN, STAT3, TNF, TP53, etc. Enrichment analysis revealed involvement of PI3K-Akt, HIF-1, FoxO, MAPK, and TNF signaling pathways. In addition, we found the most important active compounds bind tightly to core proteins, which were verified by molecular docking analysis. The AKT-related pathway had good binding energy, and the pathway was verified by cell and animal experiments. CONCLUSION: The potential mechanism and efficacy of Narenmandula against PMOP may be related to the PI3K-AKT pathway.

Urolithin A attenuates RANKL-induced osteoclastogenesis by co-regulating the p38 MAPK and Nrf2 signaling pathway.

As a critical regulator of bone resorption. osteoclastogenesis is closely associated with osteoporosis (OP) and commonly induced by receptor activator of nuclear factor-κB ligand (RANKL), suggesting that suppression of inflammation may improve OP. Urolithin A (UroA), an active metabolite of ellagic acid, is known to exert anti-inflammatory and antioxidative effects. However, whether UroA attenuates osteoclastogenesis remains unclear. Using a lipopolysaccharide (LPS)-induced bone loss model, we evaluated the effects of UroA on inflammatory osteoclastogenesis in mice and explored the potential mechanism from RANKL-related signaling pathway. UroA significantly improved LPS-induced bone loss and rescued the imbalance in bone microarchitecture parameters. Hematoxylin&eosin (H&E) and tartrate resistant acid phosphatase (TRAP) staining of femurs showed that UroA suppressed LPS-induced osteoclastogenesis accompanied by the activation of nuclear factor-erythroid 2-related factor 2 (Nrf2) signaling. In RANKL-triggered mouse bone marrow-derived macrophages (BMDMs), UroA inhibited the formation of osteoclasts and Fibrous actin rings (F-actin rings), and decreased TRAP activity. Moreover, UroA significantly decreased mRNA and protein expression of major inflammatory cytokines in LPS-challenged RAW264.7 cells by decreasing the phosphorylation of NF-κB p65, c-Jun N-terminal kinase (JNK), extracellular signal regulated kinase1/2 (Erk1/2), and p38. Furthermore, UroA may activate the Nrf2 signaling pathway by increasing mRNA and protein expression of antioxidant proteins. We conclude that UroA attenuated RANKL-induced osteoclastogenesis by suppressing the p38 mitogen-activated protein kinase (MAPK) pathway and inducing Nrf2 nuclear translocation. Thus, supplementation with UroA may help alleviate inflammation-induced bone loss and bone resorption.

Primary observation of the role of posttranslational modification of dentin sialophosphoprotein (DSPP) on postnatal development of mandibular condyle in mice.

OBJECTIVES: We aimed to observe the posttranslational role of dentin sialophosphoprotein (DSPP) on postnatal development of mandibular condyle in mice. METHODS: To explore the function of full-length DSPP, four groups of mice were employed: (1) wild type (WT) mice; (2)Dspp knockout (Dspp KO) mice; (3) mice expressing the normal DSPP transgene in the Dspp KO background (Dspp KO/normal Tg); (4) mice expressing the uncleavable full-length DSPP in the Dspp KO background (Dspp KO/D452A Tg). Firstly, Plain X-ray Radiography and Micro-computed Tomography were used to observe the condylar morphology changes of Dspp KO/D452A Tg mice in comparison with the other three groups. Then, Hematoxylin & eosin and toluidine blue staining were applied to uncover the histological changes of mandibular condylar cartilage (MCC) of Dspp KO/D452A Tg mice. To explore the function of the NH2-terminal fragments (i.e. DSP/DSP-PG), three groups of mice were employed: (1) WT mice; (2) Dspp KO mice; (3) mice expressing the NH2-terminal fragments of DSPP in the Dspp-null background (Dspp KO/DSP Tg). The former strategies were utilized to examine the differences of condylar morphology and histological structures changes within three groups of mice. RESULTS: Transgenic full-length DSPP partially maintained mandibular condylar morphology and MCC thickness of Dspp KO mice. Transgenic DSP failed to do so, but led to smaller mandibular condyle and disordered cartilage structure. CONCLUSIONS: Our observations provide insight into the role of posttranslational modification of DSPP in the postnatal development of healthy MCC and maintenance of condylar morphology.

The Interaction Between Intracellular Energy Metabolism and Signaling Pathways During Osteogenesis.

Osteoblasts primarily mediate bone formation, maintain bone structure, and regulate bone mineralization, which plays an important role in bone remodeling. In the past decades, the roles of cytokines, signaling proteins, and transcription factors in osteoblasts have been widely studied. However, whether the energy metabolism of cells can be regulated by these factors to affect the differentiation and functioning of osteoblasts has not been explored in depth. In addition, the signaling and energy metabolism pathways are not independent but closely connected. Although energy metabolism is mediated by signaling pathways, some intermediates of energy metabolism can participate in protein post-translational modification. The content of intermediates, such as acetyl coenzyme A (acetyl CoA) and uridine diphosphate N-acetylglucosamine (UDP-N-acetylglucosamine), determines the degree of acetylation and glycosylation in terms of the availability of energy-producing substrates. The utilization of intracellular metabolic resources and cell survival, proliferation, and differentiation are all related to the integration of metabolic and signaling pathways. In this paper, the interaction between the energy metabolism pathway and osteogenic signaling pathway in osteoblasts and bone marrow mesenchymal stem cells (BMSCs) will be discussed.

Haploinsufficiency of Dspp Gene Causes Dentin Dysplasia Type II in Mice.

Dentin dysplasia (DD) and dentinogenesis imperfecta (DGI) patients have abnormal structure, morphology, and function of dentin. DD-II, DGI-II, and DGI-III are caused by heterozygous mutations in the dentin sialophosphoprotein (DSPP) gene in humans. Evidences have shown that loss of function of DSPP in Dspp knockout mice leads to phenotypes similar to DGI-III, and that the abnormal dentinogenesis is associated with decreased levels of DSPP, indicating that DSPP haploinsufficiency may play a role in dentinogenesis. Thus, to testify the haploinsufficiency of Dspp, we used a Dspp heterozygous mouse model to observe the phenotypes in the teeth and the surrounding tissues. We found that Dspp heterozygous mice displayed dentin phenotypes similar to DD-II at the ages of 12 and 18 months, which was characterized by excessive attrition of the enamel at the occlusal surfaces, thicker floor dentin of the pulp chamber, decreased pulp volume, and compromised mineralization of the dentin. In addition, the periodontium was also affected, exhibiting apical proliferation of the junctional epithelium, decreased height and width of the alveolar bone, and infiltration of the inflammatory cells, leading to the destruction of the periodontium. Both the dental and periodontal phenotypes were age-dependent, which were more severe at 18 months old than those at 12 months old. Our report is the first to claim the haploinsufficiency of Dspp gene and a DD-II mouse model, which can be further used to study the molecular mechanisms of DD-II.